Preparation and characterization of a polyclonal antibody from rabbit for detection of trinitrotoluene by a surface plasmon resonance biosensor

Matsumoto, Kiyoshi; Torimaru, Akira; Ishitobi, Sachiko; Sakai, Takahisa; Ishikawa, Hiroya; Toko, Kiyoshi; Miura, Norio; Imato, Toshihiko

doi:10.1016/j.talanta.2005.08.054

Cited by 57 publications

(39 citation statements)

References 12 publications

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“…For detecting clenbuterol, the indirect competitive inhibition immunoassay technique was employed [13,[35][36][37]. In this study, the antibody solution (PBS solution containing 1 µg mL -1 of Ab) was premixed with a sample solution (PBS solution containing clenbuterol) before injection into the sensing system.…”

Section: Immunoassaysmentioning

confidence: 99%

Effect of alkanethiol molecular structure on sensitivity of surface plasmon resonance sensor

Suherman

Morita

Kawaguchi

2015

Sensors and Actuators B: Chemical

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This work aims to find the key factor determining the detection limit for an indirect competitive inhibition immunoassay using surface plasmon resonance (SPR) sensing. In our previous work, the thiol solution concentration used in a self-assembly process highly affected the alkanethiol monolayer structure on the sensor surface [Suherman et al., 2014]. It was noticed that the monolayer structure determined the immunoassay sensitivity due to the orientation and the surface concentration of antigen in domain structure of monolayer. To study the effect of orientation of antigen, we examined here three types of alkanethiol compounds: dithiobis(succinimidyl undecanoate) (DSU; straight-chain alkanethiol), carboxy-EG 6 -undecanethiol (CEG 6 ; flexible-chain alkanethiol), and(C 2 -NTA; three-branched alkanethiol). By electrochemical reductive desorption, the surface concentration of DSU, CEG 6 , and C 2 -NTA were estimated to be 6.6 × 10 -10 , 8.5 × 10 -10 , 4.6 × 10 -10 mol cm -2 , respectively. Subsequently, SPR suggested that the ratios of immobilized antigen (clenbuterol) per thiol were 0.13, 0.16, and 0.15 for DSU, CEG 6 , and C 2 -NTA, respectively. This suggests that the amount of immobilized clenbuterol was defined by the molecular size of clenbuterol. Sensor surface structures on a molecular scale were evaluated using high-resolution scanning tunneling microscopy (STM), which showed tilted clenbuterol with θ DSU > θ CEG6 > θ C2-NTA . For SPR sensing of clenbuterol, C 2 -NTA showed the highest sensitivity (LOD = 10 ppt) among the examined alkanethiols, because clenbuterol formed a laid-on structure. Based on kinetics, the key factor for sensing performance is discussed.2

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Section: Immunoassaysmentioning

confidence: 99%

Effect of alkanethiol molecular structure on sensitivity of surface plasmon resonance sensor

Suherman

Morita

Kawaguchi

2015

Sensors and Actuators B: Chemical

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“…Thus, the amount of antigen can be determined indirectly. In the inhibition assay, the limit of detection (LOD) decreases as antibody concentration decreases; (3,5) thus, it is necessary to use as low a concentration of antibody as possible for highly sensitive detection. According to the result in the previous section, the G6-dendron-based surface is the most suitable for TNT detection because it minimizes the analyte concentration required to produce a detectable signal.…”

Section: Highly Sensitive Detection Of Tnt By Inhibition Assaymentioning

confidence: 99%

“…For the past few years, we developed highly sensitive SPR immunosensors for TNT, TNP, and DNT detection using an inhibition assay. (2)(3)(4)(5)(6)(7) In these studies, hapten-protein conjugates were often used as constituent materials of SPR sensor chips (2)(3)(4)6) because the threedimensional structure of the surface-bound proteins created an easily accessible binding environment for antibodies, which were fl owed over the sensor chip surfaces, and contributed to the enhancement of SPR response.…”

Section: Introductionmentioning

confidence: 99%

Highly Sensitive Detection of TNT Using a Poly(amidoamine) Dendron-Based SPR Immunosensor

Mizuta

Onodera

Singh

et al. 2010

Sensors and Materials

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In this paper, we describe the fabrication of novel poly(amidoamine) (PAMAM) dendron-based Surface Plasmon Resonance (SPR) sensor chip surfaces supported by a self-assembled monolayer (SAM) of alkanethiol and their use in the highly sensitive detection of 2,4,6-trinitrotoluene (TNT), which is one of main components of explosives. The fabricated surfaces showed excellent binding capacity for TNT antibodies; this is probably due to the fact that the highly branched functional groups and three-dimensional structure of dendron molecules offer densely arranged binding sites and improve binding effi ciency. A limit of detection (LOD) of ca. 15 ppt (pg ml −1 ) was achieved by inhibition assay.

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“…(5)(6)(7) However, the number of binding sites of an anti-TNT antibody cannot be increased anymore, because the binding sites are arranged in two dimensions on the SPR sensor surface. Therefore, a dendron-modified SPR sensor surface was fabricated for increasing the number of binding sites -the binding sites were arranged in three dimensions.…”

Section: Introductionmentioning

confidence: 99%

Highly Sensitive Detection of 2,4,6-Trinitrotoluene (TNT) Using Poly(vinylamine-co-N-vinylformamide)-Based Surface Plasmon Resonance (SPR) Immunosensor

Yatabe¹,

Onodera

Toko

2013

Sensors and Materials

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In this paper, we describe the fabrication of a novel poly-(vinylamine-co-Nvinylformamide) (poly-(VAm-co-NVF))-based surface plasmon resonance (SPR) sensor chip supported by a self-assembled monolayer (SAM) of alkanethiol and its use in the highly sensitive detection of 2,4,6-trinitrotoluene (TNT), which is a typical explosive. 2,4-dinitrophenyl-glycine (DNP-Gly) was immobilized to amino groups on the side chain of poly-vinylamine (poly-VAm) on the sensor surface. The fabricated surface showed a high response for flow of anti-TNT antibodies. However, it was found that the response included the nonspecific adsorption of anti-TNT antibodies, because unreacted amino groups of the poly-VAm were positively charged and it caused nonspecific adsorption due to electrostatic interaction. Then, the number of amino groups was reduced by making low-hydrolyzed poly-N-vinylformamide(poly-NVF), i.e., poly(VAm-co-NVF) -poly-NVF is a precursor of poly-VAm, and nonspecific adsorption was controlled. The limit of detection (LOD) of TNT was 28 ppt using the sensor chip fabricated with 23% hydrolyzed poly-(VAm-co-NVF) by inhibition assay.

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Preparation and characterization of a polyclonal antibody from rabbit for detection of trinitrotoluene by a surface plasmon resonance biosensor

Cited by 57 publications

References 12 publications

Effect of alkanethiol molecular structure on sensitivity of surface plasmon resonance sensor

Effect of alkanethiol molecular structure on sensitivity of surface plasmon resonance sensor

Highly Sensitive Detection of TNT Using a Poly(amidoamine) Dendron-Based SPR Immunosensor

Highly Sensitive Detection of 2,4,6-Trinitrotoluene (TNT) Using Poly(vinylamine-co-N-vinylformamide)-Based Surface Plasmon Resonance (SPR) Immunosensor

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